Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines.

Gamm Lab // Publications // Dec 21 2018

PubMed ID: 30567931

Author(s): Capowski EE, Samimi K, Mayerl SJ, Phillips MJ, Pinilla I, Howden SE, Saha J, Jansen AD, Edwards KL, Jager LD, Barlow K, Valiauga R, Erlichman Z, Hagstrom A, Sinha D, Sluch VM, Chamling X, Zack DJ, Skala MC, Gamm DM. Reproducibility and staging of 3D human retinal organoids across multiple pluripotent stem cell lines. Development. 2018 Dec 19. pii: dev.171686. doi: 10.1242/dev.171686. [Epub ahead of print]

Journal: Development (Cambridge, England), Dec 2018

Numerous protocols have been described that produce neural retina from human pluripotent stem cells (hPSCs), many of which are based on the culture of 3D organoids. While nearly all such methods yield at least partial segments of highly mature-appearing retinal structure, variabilities exist within and between organoids that can change over a protracted time course of differentiation. Adding to this complexity are potential differences in the composition and configuration of retinal organoids when viewed across multiple differentiations and hPSC lines. In an effort to better understand the current capabilities and limitations of these cultures, we generated retinal organoids from 16 hPSC lines and monitored their appearance and structural organization over time by light microscopy, immunocytochemistry, metabolic imaging, and electron microscopy. We also employed optical coherence tomography and 3D imaging techniques to assess and compare whole or broad regions of organoids to avoid selection bias. Results from this study led to the development of a practical staging system to reduce inconsistencies in retinal organoid cultures and increase rigor when utilizing them in developmental studies, disease modeling and transplantation.

© 2018. Published by The Company of Biologists Ltd.